LOCUS Exported 4760 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gene deletion in S. cerevisiae and S. pombe. ACCESSION . VERSION . KEYWORDS pFA6a-LEU2MX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4760) AUTHORS Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E TITLE New vectors for epitope tagging and gene disruption in Schizosaccharomyces pombe. JOURNAL Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100. PUBMED 24215641 REFERENCE 2 (bases 1 to 4760) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4760 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 334..741 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 742..1836 /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" primer_bind complement(2397..2416) /label=T7 /note="T7 promoter, forward primer" promoter complement(2398..2416) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2503..2520) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2674..3262) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2754..2773) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3433..4293) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4056..4075 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4294..4398) /gene="bla" /label=AmpR promoter primer_bind 4466..4484 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4522..4544) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4644..4663 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4744..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4744..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta 61 aggcgcgcca gatcttgcag gcatgcaagc ttgtcgagga gaacttctag tatatctaca 121 tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa aatccacatt 181 ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt 241 ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga gcggtctaag 301 gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca ggtatcgtaa 361 gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata acgagaacac 421 acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga 481 ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga 541 gccgcggaac cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca 601 caatacttga agttgacaat attatttaag gacctattgt tttttccaat aggtggttag 661 caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata tatatatata 721 tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac 781 cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt 841 cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct 901 acaggtgtcc cacttccaga tgaggcgctg gaagcctcca agaaggttga tgccgttttg 961 ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta 1021 ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc 1081 gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt 1141 gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat 1201 ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg 1261 gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct 1321 aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa 1381 ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag 1441 aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc 1501 gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct 1561 ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat 1621 ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa 1681 ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg 1741 gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccacgga agtcggtgat 1801 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat 1861 ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga 1921 atatgttcat agggtagacg aaactatata cgcaatctac atacatttat caagaaggag 1981 aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata 2041 aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag 2101 ttaggtgtaa cagaaaatca tgaaactatg attcctaatt tatatattgg aggattttct 2161 ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg acctgaccat ttgatggagt 2221 ttaagtcaat accttcttga accatttccc ataatggtga aagttccctc aagaatttta 2281 ctctgtcaga aacggcctta acgacgtagt cgatcgacaa gctgtttaaa cgagctcgaa 2341 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct 2401 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg 2461 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 2521 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 2581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 2641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 2701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 2881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 3001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 3061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 3121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 3901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 3961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 4441 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg 4501 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 4561 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 4621 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat 4681 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat 4741 acgatttagg tgacactata //